Metabolite Analysis And Stachyose Metabolism Regulation Of Phallus Dongsun In Liquid Submerged Culture

Phallus dongsun has high edible and medicinal value,but there is no study on the composition and relationship of metabolites in its mycelium fermentation culture.In order to understand the composition of extracellular metabolites of Phallus dongsun,we analyzed the difference of mycelium secretion and its relationship at different fermentation time by submerged fermentation technology.According to the results of bioinformatics analysis,we determined the extracellular component markers of Phallus dongsun and speculated its KEGG metabolic pathway.After that,we analyzed the metabolic pathway from the aspects of substrate metabolic regulation and transcription factor expression,which provided a reference for explaining the regulation of the metabolic pathway of Phallus dongsun polysaccharide.The results are as follows:1.Identification of strains.The sequencing results of TA clone showed that all the peaks were single peak and clear,independent and complete.The total base length of the strain was 594bp,and the GC content was 51%.The results showed that the strain P1710141 was Phallus dongsun.2.Analysis of metabolites in different fermentation stages.The results of UHPLC-MS/MS analysis showed that 534 metabolites were identified,and the proportion of organic acids was17.978%,which was the main reason for the acidity of fermentation environment.Thirty-seven characteristic metabolites were identified,daidzein,stachyose,trans-4-hydroxy-L-proline and UDP Glc NAc and so on.The differential metabolites were significantly enriched in 86 metabolic pathways,including ABC transporters,c AMP signaling pathway and galactose metabolism.We speculated the synthesis pathway of exopolysaccharides such as stachyose,which has a guiding significance for the study of exopolysaccharides of phallus.In addition,the type and quantity of metabolites in different fermentation time were much higher than the chemical composition of fruiting body and mycelium,which had been reported.It has guiding significance for the multi angle study of bioactive components of Phallus.3.Objective to establish a method for the determination of carbohydrate metabolites.A HPLC-ELSD method for the determination of carbohydrate metabolites in Phallus dongsun fermentation broth was established.Asahipak NH2P-50 4E was used as the chromatographic column,the mobile phase was acetonitrile and water(62:38)and the injection volume was 3?L.The column temperature is set at 40?,the gain is 20 times,the pressure is 0.5MPa,the drift tube temperature is 60?,and the optimal flow rate is 0.5ml?min^-1.4.Analysis of metabolite transcriptional group.The effective length of logarithmic phase,stable phase and autolysis phase transcripts were 6.57G,5.78G and 5.84G,and 27426 unigenes were annotated.Association analysis of differential genes and metabolites showed that amino acid biosynthesis and lipolysis metabolism of adipocytes were significantly enriched.Aromatic and amino acid family metabolism were the reasons for the change of bacterial flavor during Phallus dongsun fermentation.According to the changes of camp and DL lactic acid content from pre logarithmic stage to post logarithmic stage,it is speculated that RP-S6e and CCNB1 are the main regulators of logarithmic phase transition in Dictyophora indusiana mycelia fermentation,and MHC-Ib class plays an inhibitory role in cell metabolism during autolysis phase of Phallus dongsun fermentation.5.The metabolic pathway of stachyose was analyzed.The analysis of stachyose metabolic pathway showed that raffinose and sucrose could be transformed into stachyose,and the expression of stachyose was inhibited when raffinose concentration was 600 mg/L,but sucrose concentration of 15g/L promoted stachyose expression,stachyose was mainly synthesized in logarithmic growth period.GLA is a key gene regulating stachyose metabolism,and its expression is up-regulated,?-Galactosidase catalyzed the hydrolysis of stachyose to raffinose and glucose.Stachyose can be hydrolyzed to produce energy through msmk and glucose through aglk,which can realize the transmembrane transport of mycelium cells.The biosynthetic pathway of stachyose in Phallus dongsun was elucidated from two aspects of substrate and transcription factor.